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Involvement of Seladin-1 in goniothalamin-induced apoptosis in urinary bladder cancer cells.

Yen HK, Fauzi AR, Din LB, McKelvey-Martin VJ, Meng CK, Inayat-Hussain SH, Rajab NF - BMC Complement Altern Med (2014)

Bottom Line: This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells.The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Science Programme, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia. nfadilah@medic.ukm.my.

ABSTRACT

Background: Selective Alzheimer Disease Indicator-1 (or Seladin-1) is a multifunctional protein first discovered by downregulation of its expression in Alzheimer's disease. Interestingly, the expression of this protein is upregulated in several cancers, including primary bladder cancer. However, its role in cancer formation has yet to be discovered. Goniothalamin is a natural product that has been demonstrated to induce apoptosis in various cancer cell lines. In this study, we have elucidated the role of Seladin-1 in goniothalamin-induced cytotoxicity towards human urinary bladder cancer cell line RT4.

Methods: The cytotoxicity of goniothalamin in human urinary bladder cancer cell line RT4 was assessed using MTT assay and the mode of cell death was determined by Annexin V-FITC/PI labeling assay. Finally, the expression of Seladin-1 protein in goniothalamin-treated RT4 cells was determined by Western blot.

Results: MTT assay showed that the cytotoxicity of goniothalamin on RT4 cells was concentration and time dependent with IC50 values of 61 μM (24 hr), 38 μM (48 hr) and 31 μM for 72 hr, respectively. Cell death induced was confirmed through apoptosis; as assessed using the Annexin V-FITC/PI labeling assay. Furthermore, the involvement of Seladin-1 in goniothalamin-induced apoptosis was evidenced through the cleavage of 60 kDa protein to 40 kDa and 20 kDa. This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.

Conclusion: This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells. The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

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Related in: MedlinePlus

Percentage of viability of RT4 cells treated by etoposide (18.75, 37.5, 75, 150, and 300 μM) for 24, 48, and 72 hrs. Each point represents mean ± SEM of 3 different experiments (*p < 0.05 compared to the control).
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Fig2: Percentage of viability of RT4 cells treated by etoposide (18.75, 37.5, 75, 150, and 300 μM) for 24, 48, and 72 hrs. Each point represents mean ± SEM of 3 different experiments (*p < 0.05 compared to the control).

Mentions: The cytotoxic effect of goniothalamin on RT4 cells was assessed by MTT assay. Following treatment of 24, 48 and 72 hr, the percentage viability of the cells was obtained and graph was plotted versus concentration of goniothalamin (see Figure 1). The graph shows that the IC50 value was 61 μM, 38 μM and 31 μM following 24 hrs, 48 hrs and 72 hrs treatment, respectively. The viability drop from 100% from control and further reduced across concentrations of goniothalamin and time points; with the highest concentration (100 μM) of goniothalamin showed less than 20% of cells were viable following 24 hrs, 48 hrs, and 72 hrs of treatment. Statistical analysis showed that there was a significant reduction in the percentage of cell viability following treatment with goniothalamin at each concentration (i.e., 6.25, 12.5, 25, 50, and 100 μM) compared to control or untreated cells (p < 0.05). Etoposide was used as a positive control for this experiment and the results of the cytotoxic effect are shown in Figure 2. The IC50 value of etoposide on RT4 cells was 46 μM, 15 μM and 14 μM following 24 hrs, 48 hrs and 72 hrs treatment, respectively. There was a significant reduction in percent viability compared to the control at each concentration of etoposide (p < 0.05).Figure 1


Involvement of Seladin-1 in goniothalamin-induced apoptosis in urinary bladder cancer cells.

Yen HK, Fauzi AR, Din LB, McKelvey-Martin VJ, Meng CK, Inayat-Hussain SH, Rajab NF - BMC Complement Altern Med (2014)

Percentage of viability of RT4 cells treated by etoposide (18.75, 37.5, 75, 150, and 300 μM) for 24, 48, and 72 hrs. Each point represents mean ± SEM of 3 different experiments (*p < 0.05 compared to the control).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4150971&req=5

Fig2: Percentage of viability of RT4 cells treated by etoposide (18.75, 37.5, 75, 150, and 300 μM) for 24, 48, and 72 hrs. Each point represents mean ± SEM of 3 different experiments (*p < 0.05 compared to the control).
Mentions: The cytotoxic effect of goniothalamin on RT4 cells was assessed by MTT assay. Following treatment of 24, 48 and 72 hr, the percentage viability of the cells was obtained and graph was plotted versus concentration of goniothalamin (see Figure 1). The graph shows that the IC50 value was 61 μM, 38 μM and 31 μM following 24 hrs, 48 hrs and 72 hrs treatment, respectively. The viability drop from 100% from control and further reduced across concentrations of goniothalamin and time points; with the highest concentration (100 μM) of goniothalamin showed less than 20% of cells were viable following 24 hrs, 48 hrs, and 72 hrs of treatment. Statistical analysis showed that there was a significant reduction in the percentage of cell viability following treatment with goniothalamin at each concentration (i.e., 6.25, 12.5, 25, 50, and 100 μM) compared to control or untreated cells (p < 0.05). Etoposide was used as a positive control for this experiment and the results of the cytotoxic effect are shown in Figure 2. The IC50 value of etoposide on RT4 cells was 46 μM, 15 μM and 14 μM following 24 hrs, 48 hrs and 72 hrs treatment, respectively. There was a significant reduction in percent viability compared to the control at each concentration of etoposide (p < 0.05).Figure 1

Bottom Line: This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells.The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Science Programme, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia. nfadilah@medic.ukm.my.

ABSTRACT

Background: Selective Alzheimer Disease Indicator-1 (or Seladin-1) is a multifunctional protein first discovered by downregulation of its expression in Alzheimer's disease. Interestingly, the expression of this protein is upregulated in several cancers, including primary bladder cancer. However, its role in cancer formation has yet to be discovered. Goniothalamin is a natural product that has been demonstrated to induce apoptosis in various cancer cell lines. In this study, we have elucidated the role of Seladin-1 in goniothalamin-induced cytotoxicity towards human urinary bladder cancer cell line RT4.

Methods: The cytotoxicity of goniothalamin in human urinary bladder cancer cell line RT4 was assessed using MTT assay and the mode of cell death was determined by Annexin V-FITC/PI labeling assay. Finally, the expression of Seladin-1 protein in goniothalamin-treated RT4 cells was determined by Western blot.

Results: MTT assay showed that the cytotoxicity of goniothalamin on RT4 cells was concentration and time dependent with IC50 values of 61 μM (24 hr), 38 μM (48 hr) and 31 μM for 72 hr, respectively. Cell death induced was confirmed through apoptosis; as assessed using the Annexin V-FITC/PI labeling assay. Furthermore, the involvement of Seladin-1 in goniothalamin-induced apoptosis was evidenced through the cleavage of 60 kDa protein to 40 kDa and 20 kDa. This was followed by a gradual increase of 20 kDa fragment suggesting the involvement of Seladin-1 in goniothalamin-induced apoptosis on RT4 cells.

Conclusion: This study demonstrates that goniothalamin induce cytotoxicity and apoptosis on RT4 cells. The involvement of Seladin-1 in goniothalamin-induced apoptosis further suggested that Seladin-1 may play a role in the formation of primary bladder cancer.

Show MeSH
Related in: MedlinePlus